1.Construction of survivin shRNA and APC double gene vector and its expression in HT-29 cells
Xixian YUAN ; Houyu DUAN ; Feng CAO ; Jingfeng DU ; Yanli ZHU ; Kai CHENG ; Yantao YANG
Journal of Medical Postgraduates 2016;29(4):369-374
Objective Studies show that the abnormal ex-pressions of APC and survivin play important roles in the development and progression of colon cancer .Survivin shRNA and APC double gene co-expression lentiviral vector was constructed to observe whether it could be successfully expressed in HT-29 colon cancer cells and whether it could impact on colon cancer cell apoptosis . Methods We selected the best shRNA interference fragment from the construc-tion of three pairs of complementary shRNA fragments and connected it with the effective fragment of APC ( aa1020-1698 ) to construct a double gene co-expression lentiviral vector .HT-29 cells were divid-ed into experimental group , empty loading group and blank group .HT-29 cells were observed by fluorescence microscopy after infec-tion.Survivin and APC expression levels were observed by real time PCR and western blot .Apoptosis was detected by caspase-3 activi-ty assay. Results ①We successfully constructed three pairs of shRNA sequences and proved that they had no human gene homolo -gous to the rest.Real time PCR analysis showed that the best sequence was shRNA 3.②After the sequence alignment of constructed shRNA vectors, three pairs of shRNA sequences were completely the same with the first designed sequence .Green fluorescence was observed in HT-29 cells by fluorescence microscope .The survivin content in experiment group (31.71 ±1.49) was significantly de-creased compared with empty loading group (100 ±0) and blank group(95.12 ±2.15)(P<0.05).The expression level of APC mR-NA was significantly increased compared with empty loading group (0 ±0) and blank group(0.51 ±0.15)(P<0.05).③The relative ratio of apoptosis in experiment group (0.573 ±0.050) was significantly increased compared with empty loading group (0.390 ± 0.040) and blank group(0.407 ±0.030)(P<0.05). Conclusion We have successfully constructed survivin-shRNA-APC double gene co-expression lentiviral vector which can be successfully expressed in HT-29 colon cancer cells , providing references for subse-quent gene therapy by the use of the carrier .
2.Study on correlations between total saponins content in rhizome or mycorrhizal infection rate of Pairs polyphylla var. yunnanensis and soil factors.
Yantao DUAN ; Zhongjun HE ; Shewang LIANG ; Yin WANG ; Junfen XIONG ; Fangdong ZHAN ; Rui ZENG
China Journal of Chinese Materia Medica 2011;36(22):3091-3095
OBJECTIVEThrough correlation and path analysis between total saponins content in rhizome/mycorrhizal infection rate in roots of Pairs polyphylla var. yunnanensis and soil factors, to make an inquiry into the role of soil factors in the quality formation of P. polyphylla var. yunnanensis.
METHODTested total saponins in rhizome, mycorrhizal fungal infection rate in root and physical and chemical properties in rhizosphere soil in 25 different growth areas, and statistically analyzed the relationship between total saponins in rhizome/mycorrhizal infection rate in roots of P. polyphylla var. yunnanensis and soil factors by using correlation and path analysis.
RESULTThe symbiosis relationship between AM mycorrhizal and roots of P. polyphylla var. yunnanensis were better established under natural condition, of which the infection ratio between 36.41%, 83.37%. There were significantly positive correlation between total saponins content in rhizome and urease activity or alkaline phosphatase activities or organic matter in soil, but there was significantly negative correlation between total saponins content and bulk density. There was significantly positive correlation between AM infection ratio and alkaline nitrogen. Path analysis indicated that total saponins of rhizome mainly affected by alkaline nitrogen in soil rhizosphere, secondly by soil organic matter and soil urease activity. While the mycorrhizal fungal colonization ratio was mainly affected by soil pH, secondly by alkaline nitrogen, urease activity, and available phospherus in soil.
CONCLUSIONThere is closed relationship between quality formation of P. polyphylla var. yunnanensis and soil factors. Path analysis is better for reflecting the contribution of soil factors to total saponins and mycorrhizal infection ratio.
Alkaline Phosphatase ; metabolism ; Liliaceae ; chemistry ; microbiology ; Mycorrhizae ; growth & development ; Rhizome ; chemistry ; Saponins ; analysis ; Soil ; analysis ; Urease ; metabolism
3.Sensing of nutrients and intestinal microecology
Chinese Journal of Clinical Nutrition 2019;27(3):188-192
Sensing of nutrients and intestinal microecology are necessary for life.The nutrient sensing molecules are key factors to regulate a series of pathophysiological processes in nutrition metabolism.Microbial sensing molecules in the gut play critical roles in intestinal microecology and maintenance of the gut homeostasis.However,the specific mechanism remains to be further explored.This article reviews the intestinal nutrient sensing system and the molecular regulation mechanism of the intestinal microelogy.
4.Effect of previous intestinal resection on postoperative anastomotic fistula in Crohn′s disease with intestinal resection and anastomosis
Yantao DUAN ; Yuhua HUANG ; Bin LIU ; Danhua YAO ; Lei ZHENG ; Zhiyuan ZHOU ; Pengfei WANG ; Yousheng LI
Chinese Journal of General Surgery 2020;35(8):620-623
Objective:To explore the effect of previous intestinal resection on anastomotic fistula within 30 days after surgery in Crohn′s disease.Methods:The clinical data from 92 Crohn′s disease patients who underwent intestinal resection and anastomosis at the Department of General Surgery in Shanghai Ninth People′s Hospital, Shanghai Jiaotong University School of Medicine from Jan 2016 to Sep 2019. Patients were divided into no previous intestinal resection group ( n=45) and previous intestinal resection group ( n=47). The relationship between previous intestinal resection and postoperative anastomotic leak in Crohn′s disease patients with intestinal resection and anastomosis was analyzed. Results:A total of 11 cases (12% leak rate) underwent postoperative anastomotic leak. There were 2 leaks in patients with no previous history of intestinal resection, while 9 leaks in patients with previous bowel resection (χ 2 =4.722, P=0.03). The OR of the postoperative anastomotic leak in Crohn′s disease patients with previous intestinal resection compared with no previous intestinal resection group was 5.092 (95% CI: 1.035-25.048). Patients with 1 previous resection (24 cases) had a leak rate of 13%, whereas patients with >1 previous resection episodes (23 cases) had a leak rate of 26%. The number of previous resection episodes correlated with an increasing risk for clinical anastomotic leak (correlation coefficien r=0.995). Conclusions:Previous intestinal resection is an independent risk factor for ensuing postoperative anastomotic leak in Crohn′s disease patients with intestinal resection and anastomosis.
5.Invasiveness assessment by CT quantitative and qualitative features of lung cancers manifesting ground-glass nodules in 555 patients: A retrospective cohort study
Yantao YANG ; Wei WANG ; Yichen YANG ; Biying WANG ; Huilian HU ; Ziqi JIANG ; Dezhong CAI ; Yaowu DUAN ; Jiezhi JIANG ; Jia LUO ; Guangqiang ZHAO ; Yunchao HUANG ; Lianhua YE
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2024;31(01):51-58
Objective To explore the correlation between the quantitative and qualitative features of CT images and the invasiveness of pulmonary ground-glass nodules, providing reference value for preoperative planning of patients with ground-glass nodules. Methods The patients with ground-glass nodules who underwent surgical treatment and were diagnosed with pulmonary adenocarcinoma from September 2020 to July 2022 at the Third Affiliated Hospital of Kunming Medical University were collected. Based on the pathological diagnosis results, they were divided into two groups: a non-invasive adenocarcinoma group with in situ and minimally invasive adenocarcinoma, and an invasive adenocarcinoma group. Imaging features were collected, and a univariate logistic regression analysis was conducted on the clinical and imaging data of the patients. Variables with statistical difference were selected for multivariate logistic regression analysis to establish a predictive model of invasive adenocarcinoma based on independent risk factors. Finally, the sensitivity and specificity were calculated based on the Youden index. Results A total of 555 patients were collected. The were 310 patients in the non-invasive adenocarcinoma group, including 235 females and 75 males, with a meadian age of 49 (43, 58) years, and 245 patients in the invasive adenocarcinoma group, including 163 females and 82 males, with a meadian age of 53 (46, 61) years. The binary logistic regression analysis showed that the maximum diameter (OR=4.707, 95%CI 2.060 to 10.758), consolidation/tumor ratio (CTR, OR=1.027, 95%CI 1.011 to 1.043), maximum CT value (OR=1.025, 95%CI 1.004 to 1.047), mean CT value (OR=1.035, 95%CI 1.008 to 1.063), spiculation sign (OR=2.055, 95%CI 1.148 to 3.679), and vascular convergence sign (OR=2.508, 95%CI 1.345 to 4.676) were independent risk factors for the occurrence of invasive adenocarcinoma (P<0.05). Based on the independent predictive factors, a predictive model of invasive adenocarcinoma was constructed. The formula for the model prediction was: Logit(P)=–1.293+1.549×maximum diameter of lesion+0.026×CTR+0.025×maximum CT value+0.034×mean CT value+0.72×spiculation sign+0.919×vascular convergence sign. The area under the receiver operating characteristic curve of the model was 0.910 (95%CI 0.885 to 0.934), indicating that the model had good discrimination ability. The calibration curve showed that the predictive model had good calibration, and the decision analysis curve showed that the model had good clinical utility. Conclusion The predictive model combining quantitative and qualitative features of CT has a good predictive ability for the invasiveness of ground-glass nodules. Its predictive performance is higher than any single indicator.